Abstract: The bud of Anoectochilus roxburghii(wall.)lindl. Huaye was used to to study the effects of the key factors such as culture medium(i.e., MS, B5, improved MS), sugar concentration and ipperon concentration on the plant growth control and pollution control. The suitable medium formulation was selected in order to establish test tube flowers technology of Anoectochilus roxburghii(wall.)lindl. Huaye.The results showed that the improved MS medium with sugar 20 g·L^-1 was beneficial to the plant growth control of Anoectochilus roxburghii(wall.)lindl. Huaye; ipperon 1.5 mg·L^-1 was added to the medium, which was beneficial to the pollution control of the medium; that is to say, the medium for the test-tube flowering of Anoectochilus roxburghii(wall.)lindl. Huaye was composed of improved MS medium, sugar 20 g·L^-1, ipperon 1.5 mg·L^-1, agar powder 6.0 g·L^-1, and edible pigment 0.02 g·L^-1. The plant growth control and the ornamental quality was very good in 6 months. It provides a technical basis for the development and application of test tube flowers of Anoectochilus roxburghii(wall.)lindl. Huaye.